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111.
Diran Herebian Ute Spiekerkötter Marc Lamshöft Eva Thimm Maurice Laryea Ertan Mayatepek 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》2009,877(14-15):1453-1459
In this study, we describe a bioanalytical method for quantification of NTBC in plasma of patients with hereditary tyrosinemia type 1 (HT-1) using high-performance liquid chromatography coupled to tandem mass spectrometry (LC–MS/MS). After protein precipitation with acetonitrile including Mesotrione as internal standard, separation of NTBC was achieved by RP-HPLC. Detection was performed by positive ion electrospray ionization (ESI) in selected reaction monitoring (SRM) mode. NTBC recovery in the developed method was found to be more than 90%. The lower limit of quantification was calculated to be 0.35 μM. The intra-day and inter-day precision of three different quality control samples (measured as RSD%) was less than 10% and 15%, respectively. The standard calibration curves showed good linearity within the range of 2.5–40 μM and the determined correlation coefficients were r2 ≥ 0.995. This presented method is rapid, sensitive, specific and suitable for clinical practice and research. 相似文献
112.
Killing of virally infected cells or tumor cells by cytotoxic T lymphocytes requires targeting of lytic granules to the junction between the CTL and its target. We used whole-cell patch clamp to measure the cell capacitance at fixed intracellular [Ca2+] to study fusion of lytic granules in human CTLs. Expression of a fluorescently labeled human granzyme B construct allowed identification of lytic granule fusion using total internal reflection fluorescence microscopy. In this way capacitance steps due to lytic granule fusion were identified. Our goal was to determine the size of fusing lytic granules and to describe their behavior at the plasma membrane. On average, 5.02 ± 3.09 (mean ± s.d.) lytic granules were released per CTL. The amplitude of lytic granule fusion events was ~ 3.3 fF consistent with a diameter of about 325 nm. Fusion latency was biphasic with time constants of 15.9 and 106 seconds. The dwell time of fusing lytic granules was exponentially distributed with a mean dwell time of 28.5 seconds. Fusion ended in spite of the continued presence of granules at the immune synapse. The mobility of fusing granules at the membrane was indistinguishable from that of lytic granules which failed to fuse. While dwelling at the plasma membrane lytic granules exhibit mobility consistent with docking interspersed with short periods of greater mobility. The failure of lytic granules to fuse when visible in TIRF at the membrane may indicate that a membrane-confined reaction is rate limiting. 相似文献
113.
Tanja Goyarts Klaus-Peter Brüssow Hana Valenta Ute Tiemann Kathrin Jäger Sven Dänicke 《Mycotoxin Research》2010,26(2):119-131
Six pregnant sows of 180.6 ± 5.6 kg were fed either a Fusarium-contaminated (4.42 mg DON and 48.3 μg ZON per kg, DON per os, n = 3) or a control diet (0.15 mg DON and 5 μg ZON/kg) in the period of days 63 and 70 of gestation. On day 63 of gestation,
sows fed the control diet were implanted with an intraperitoneal osmotic minipump (delivery rate of 10 μL/h, for 7 days) containing
50 mg pure (98%) DON in 2 ml 50% DMSO (DON ip, n = 3). Frequent plasma samples were taken to estimate the kinetics after oral and ip DON exposure. The intended continuous
delivery of DON by the intraperitoneal minipump could not be shown, as there was a plasma peak (Cmax) of 4.2–6.4 ng DON/mL either immediately (sow IP-2+3) or 2.5 h (sow IP-1) after implantation of the pump followed by a one-exponential
decline with a mean half-time (t1/2) of 1.75–4.0 h and only negligible DON plasma concentrations after 12 h. Therefore, the DON ip exposure has to be regarded
as one single dose 1 week before termination of experiment. The DON per os sows showed a mean basis level (after achieving
a steady state) of DON plasma concentration of about 6–8 ng/mL, as also indicated by the plasma DON concentration at the termination
of the experiment. On day 70, caesarean section was carried out, the fetuses were killed immediately after birth, and samples
of plasma, urine, and bile were taken to analyze the concentration of DON and its metabolite de-epoxy-DON. At necropsy there
were no macroscopic lesions observed in any organ of either sows or piglets. Histopathological evaluation of sows liver and
spleen revealed no alterations. The proliferation rate of peripheral blood mononuclear cells (PBMC) with or without stimulation
was not affected by the kind of DON treatment. The exposure of pregnant sows at mid-gestation (days 63–70, period of organogenesis)
to a Fusarium toxin-contaminated diet (4.42 mg DON and 0.048 mg ZON per kg) or pure DON via intraperitoneal osmotic minipump did not cause
adverse effects on health, fertility, maintenance of pregnancy, and performance of sows and their fetuses. However, DON was
detected in fetus plasma, indicating that this toxin can pass the placental barrier and may cause changes in the proportion
of white blood cells (lower monocyte and neutrophil and higher lymphocyte proportion in DON per os fetuses). 相似文献
114.
Min Zhong Wang Shen Kenneth J. Barr Jennifer P. Arbitrario Michelle R. Arkin Minna Bui Teresa Chen Brian C. Cunningham Marc J. Evanchik Emily J. Hanan Ute Hoch Karen Huen Jennifer Hyde Jeffery L. Kumer Teresa Lac Chris E. Lawrence Jose R. Martell Johan D. Oslob Kumar Paulvannan Saileta Prabhu W. Mike Flanagan 《Bioorganic & medicinal chemistry letters》2010,20(17):5269-5273
This letter describes the discovery of a novel series of tetrahydroisoquinoline (THIQ)-derived small molecules that potently inhibit both human T-cell migration and super-antigen induced T-cell activation through disruption of the binding of integrin LFA-1 to its receptor, ICAM-1. In addition to excellent in vitro potency, 6q shows good pharmacokinetic properties and its ethyl ester (6t) demonstrates good oral bioavailability in both mouse and rat. Either intravenous administration of 6q or oral administration of its ethyl ester (6t) produced a significant reduction of neutrophil migration in a thioglycollate-induced murine peritonitis model. 相似文献
115.
Studying global change through investigation of the plastic responses of xylem anatomy in tree rings 总被引:1,自引:0,他引:1
Patrick Fonti Georg von Arx Ignacio García-González Britta Eilmann Ute Sass-Klaassen Holger Gärtner Dieter Eckstein 《The New phytologist》2010,185(1):42-53
Variability in xylem anatomy is of interest to plant scientists because of the role water transport plays in plant performance and survival. Insights into plant adjustments to changing environmental conditions have mainly been obtained through structural and functional comparative studies between taxa or within taxa on contrasting sites or along environmental gradients. Yet, a gap exists regarding the study of hydraulic adjustments in response to environmental changes over the lifetimes of plants. In trees, dated tree-ring series are often exploited to reconstruct dynamics in ecological conditions, and recent work in which wood-anatomical variables have been used in dendrochronology has produced promising results. Environmental signals identified in water-conducting cells carry novel information reflecting changes in regional conditions and are mostly related to short, sub-annual intervals. Although the idea of investigating environmental signals through wood anatomical time series goes back to the 1960s, it is only recently that low-cost computerized image-analysis systems have enabled increased scientific output in this field. We believe that the study of tree-ring anatomy is emerging as a promising approach in tree biology and climate change research, particularly if complemented by physiological and ecological studies. This contribution presents the rationale, the potential, and the methodological challenges of this innovative approach. 相似文献
116.
The COP1/SPA complex and DET1 function to suppress photomorphogenesis in dark-grown Arabidopsis seedlings. Additionally, they
inhibit flowering under non-inductive short-day conditions. The COP1/SPA complex and DET1, as part of the CDD complex, represent
distinct high-molecular-weight complexes in Arabidopsis. Here, we provide genetic evidence that these complexes co-act in
regulating plant development. We report the isolation of a spa1 enhancer mutation that represents a novel, very weak allele of det1. This det1
esp1
mutation caused no detectable mutant phenotype in the presence of wild-type SPA1, but showed strongly synergistic genetic interaction with the spa1 mutation in the control of seedling photomorphogenesis, anthocyanin accumulation, plant size as well as flowering time. On
the biochemical level, the det1
esp1
spa1 double mutant showed higher HY5 protein levels than either single mutant or the wild type. The genetic interaction of spa1 and det1 mutations was further confirmed in the spa1 det1-1 double mutant which carries a strong allele of det1. Taken together, these results show that SPA1 and DET1 act together to control photomorphogenesis throughout plant development.
Hence, this suggests that COP1/SPA complexes and the CDD complex co-act in controlling the protein stability of COP1/SPA target
proteins. 相似文献
117.
Sponges (Porifera) are filter feeders that take up microorganisms from seawater and digest them by phagocytosis. At the same
time, many sponges are known to harbor massive consortia of symbiotic microorganisms, which are phylogenetically distinct
from those in seawater, within the mesohyl matrix. In the present study, feeding experiments were performed to investigate
whether phylogenetically different bacterial isolates, hereafter termed “food bacteria,” microbial seawater consortia, and
sponge symbiont consortia are taken up and processed differently by the host sponge. Aplysina aerophoba retained high numbers of bacterial isolates and microbial seawater consortia with rates of up to 2.76 × 106 bacteria (g sponge wet weight)–1 h–1, whereas the retention of sponge symbionts was lower by nearly two orders of magnitude [5.37 × 104 bacteria (g sponge wet weight)−1 h–1]. In order to visualize the processing of a food bacterium within sponge tissues, the green fluorescent protein-labeled Vibrio strain MMW1, which had originally been isolated from A. aerophoba, was constructed. Incubation of this strain with A. aerophoba and subsequent visualization in tissue cryosections showed its presence in the choanocytes and/or endopinacocytes lining
the canals but, unlike latex beads, not in deeper regions of the mesohyl, which suggests digestion of the bacteria upon contact
with the host. Denaturing gradient gel electrophoresis (DGGE) was performed on the incubation seawater to monitor the changes
in phylogenetic composition after incubation of the sponge with either seawater or sponge symbiont consortia. However, the
DGGE experiment provided no evidence for selective processing of individual lineages by the host sponge. In conclusion, this
study extends early studies by Wilkinson et al. (Proc R Soc London B 220:519–528, 1984) that sponges, here A. aerophoba, are able to differentiate between food bacteria and their own bacterial symbionts. 相似文献
118.
Schmitt S Weisz JB Lindquist N Hentschel U 《Applied and environmental microbiology》2007,73(7):2067-2078
Many marine demosponges contain large amounts of phylogenetically complex yet highly sponge-specific microbial consortia within the mesohyl matrix, but little is known about how these microorganisms are acquired by their hosts. Settlement experiments were performed with the viviparous Caribbean demosponge Ircinia felix to investigate the role of larvae in the vertical transmission of the sponge-associated microbial community. Inspections by electron microscopy revealed large amounts of morphologically diverse microorganisms in the center of I. felix larvae, while the outer rim appeared to be devoid of microorganisms. In juveniles, microorganisms were found between densely packed sponge cells. Denaturing gradient gel electrophoresis (DGGE) was performed to compare the bacterial community profiles of adults, larvae, and juvenile sponges. Adults and larvae were highly similar in DGGE band numbers and banding patterns. Larvae released by the same adult individual contained highly similar DGGE banding patterns, whereas larvae released by different adult individuals showed slightly different DGGE banding patterns. Over 200 bands were excised, sequenced, and phylogenetically analyzed. The bacterial diversity of adult I. felix and its larvae was comparably high, while juveniles showed reduced diversity. In total, 13 vertically transmitted sequence clusters, hereafter termed "IF clusters," that contained sequences from both the adult sponge and offspring (larvae and/or juveniles) were found. The IF clusters belonged to at least four different eubacterial phyla and one possibly novel eubacterial lineage. In summary, it could be shown that in I. felix, vertical transmission of microorganisms through the larvae is an important mechanism for the establishment of the sponge-microbe association. 相似文献
119.
Fieseler L Hentschel U Grozdanov L Schirmer A Wen G Platzer M Hrvatin S Butzke D Zimmermann K Piel J 《Applied and environmental microbiology》2007,73(7):2144-2155
Numerous marine sponges harbor enormous amounts of as-yet-uncultivated bacteria in their tissues. There is increasing evidence that these symbionts play an important role in the synthesis of protective metabolites, many of which are of great pharmacological interest. In this study, genes for the biosynthesis of polyketides, one of the most important classes of bioactive natural products, were systematically investigated in 20 demosponge species from different oceans. Unexpectedly, the sponge metagenomes were dominated by a ubiquitously present, evolutionarily distinct, and highly sponge-specific group of polyketide synthases (PKSs). Open reading frames resembling animal fatty acid genes were found on three corresponding DNA regions isolated from the metagenomes of Theonella swinhoei and Aplysina aerophoba. Their architecture suggests that methyl-branched fatty acids are the metabolic product. According to a phylogenetic analysis of housekeeping genes, at least one of the PKSs belongs to a bacterium of the Deinococcus-Thermus phylum. The results provide new insights into the chemistry of sponge symbionts and allow inference of a detailed phylogeny of the diverse functional PKS types present in sponge metagenomes. Based on these qualitative and quantitative data, we propose a significantly simplified strategy for the targeted isolation of biomedically relevant PKS genes from complex sponge-symbiont associations. 相似文献
120.